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引用次数: 0
摘要
L-天冬酰胺酶合成酶是一种依赖 ATP 的酶,其催化活性需要 ATP。然而,由于 ATP 的成本过高,L-天冬酰胺酶合成酶在工业生产过程中的应用受到限制。为解决这一局限性,本研究利用大肠杆菌的葡萄糖代谢与 L-天冬酰胺酶合成酶催化协同作用,探索构建高效的 ATP 再生系统。在摇瓶中确定了 L-天冬酰胺产量的最佳条件。在特定参数(包括 0.1 摩尔/升底物、0.2 摩尔/升葡萄糖、0.01 摩尔/升氯化镁,pH 值为 7.5,温度为 37 °C,搅拌速度为 300 r/min,持续 12 小时)下,L-天冬酰胺的最高产量为 2.7 克/升。放大工艺显著提高了 L-天冬酰胺的产量。经过 20 小时的转化,L-天冬酰胺的产量增加到 38.49 克/升,摩尔转化率达到 29.16%。事实证明,这种策略能有效提高 L-天冬酰胺的生产效率。与体外 ATP 再生方法相比,这种体内方法效率更高,成本更低。这些发现为酶法合成 L-天冬酰胺的工业应用提供了重要启示。
Enhancing L-asparagine Production Through In Vivo ATP Regeneration System Utilizing Glucose Metabolism of Escherichia coli.
L-asparaginase synthetase, an ATP-dependent enzyme, necessitates ATP for its catalytic activity. However, the integration of L-asparaginase synthetase into industrial processes is curtailed by the prohibitive cost of ATP. To address this limitation, this study explores the construction of an efficient ATP regeneration system using the glucose metabolism of Escherichia coli, synergistically coupled with L-asparaginase synthetase catalysis. The optimal conditions for L-asparagine yield were determined in shake flasks. A total of 2.7 g/L was the highest yield achieved under specific parameters, including 0.1 mol/L of substrate, 0.2 mol/L glucose, 0.01 mol/L MgCl2 at pH 7.5, a temperature of 37 °C, and agitation at 300 r/min over 12 h. The process was then scaled to a 3-L fermenter, optimizing the addition rates of the substrate and magnesium chloride, and employing a constant glucose feed of 10 g/L/h. The scale-up process led to a significant enhancement in the production of L-asparagine. The yield of L-asparagine was increased to 38.49 g/L after 20 h of conversion, and the molar conversion rate reached 29.16%. This strategy has proven to be effective in improving the efficiency of L-asparagine production. When compared to in vitro ATP regeneration methods, this in vivo approach showcased superior efficiency and reduced costs. These findings furnish pivotal insights that may propel the enzymatic synthesis of L-asparagine toward viable industrial application.
期刊介绍:
This journal is devoted to publishing the highest quality innovative papers in the fields of biochemistry and biotechnology. The typical focus of the journal is to report applications of novel scientific and technological breakthroughs, as well as technological subjects that are still in the proof-of-concept stage. Applied Biochemistry and Biotechnology provides a forum for case studies and practical concepts of biotechnology, utilization, including controls, statistical data analysis, problem descriptions unique to a particular application, and bioprocess economic analyses. The journal publishes reviews deemed of interest to readers, as well as book reviews, meeting and symposia notices, and news items relating to biotechnology in both the industrial and academic communities.
In addition, Applied Biochemistry and Biotechnology often publishes lists of patents and publications of special interest to readers.